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《Biotechnology Bulletin》 2011-06
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Construction of a Fosmid Library of Wuyiencin' s Producing Strain and Obtaining of the Library Probe

Tan Beibei Sun Lei Zhang Kecheng Yang Zhenjuan Wu Zhe Zhang Jun (State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,CAAS,Beijing 100193)  
Streptomyces ahygroscopicus var.wuyiensis was taken as the material.Its total genome was digested partially by enzyme Sau 3AⅠ,and 35-45 kb fragments was extracted.Then the extracted fragments were ligated to vector pCC1FOS.After packaged and transfected,the Fosmid library of Wuyiencin's producing strain was constructed.The titer of the Fosmid library was 8.8×10~5 CFU/mL. We picked 16 positive clones and digested with EcoRⅠand HindⅢ.As a result,the electrophoresis showed that the average sizes of the inserting DNA ligated to the vectors were greater than 35 kb,and the inserting rate was 100%.According to the biosynthetic gene cluster of Nikkomycin,polyoxin and macrolides antibiotics,we designed some specific primers to PCR using genome DNA as template for probe screening.The result showed that the primes designed by using PKS amplified DNA fragment about 1 693 bp which had high similarity with other biosynthetic genes of macrolides antibiotics.Construction of a Fosmid library of Wuyiencin's producing strain and obtaining of the library probe will make a foundation for cloning of biosynthetic genes of Wuyiencin.
【Fund】: 公益性行业科研专项资金项目(201103002);; 中国农业科学院植物保护研究所中央级公益性科研院所基本科研业务费专项资金项目(161014201 1004)
【CateGory Index】: S482.2
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